Fig 3.

Syn mutants are not overexpressed on the cell surface. (A) Surface expression in CHO cells of WT gB and the three syn point mutants was assessed by CELISA with the pAb R68. Nonspecific binding of R68 to cells was measured with cells transfected with pCAGGS vector instead of gB. In each experiment, the vector control was subtracted from each sample (present in triplicate). The results determined with triplicate samples were averaged, and the absorbance is expressed as a percentage of WT (100%). An average of the results of at least three experiments is reported. (B) Surface expression of WT gB, the syn point mutant A855V, and the three truncation mutants was assessed by FACS with the pAb R68. The percentage of transfected cells positive for FITC signal, indicating bound secondary antibody, was measured, and data are presented relative to WT gB (100%). (C) Overall levels of expression of WT gB and gB851 in CHO cell lysates were compared by Western blot analysis using the pAb R68. Actin was used as a loading control. MW, molecular weight (in thousands).